HIV Infections ; diagnosis ; therapy ; Humans ; Medicine, Chinese Traditional ; methods

Objective: To explore the characteristics of IR spectra of the crude drug Herba Sedi collected in different habitats and seasons.Methods: The infrared spectra of all the samples of the whole plants were obtained by FT-IR spectroscopy,the sample powder tablets were used,and identifi cation features were determined through comparative study.Results: The harvest seasons of the samples are remarkly related to the positions(cm-1),intensity or ratios of some major absorbing peaks of the infrared spectra from the 15 samples of several different months and 4 habitats in Hubei Province,China.As for peak positions,the peak values(1625?7)cm-1 of all the samples in spring are less than 1623cm-1,but in autumn(April to October) are generally more than 1628 cm-1.And peak values(1057?16)cm-1 of the samples of spring are more than 1050 cm-1,but less than 1050 cm-1 for the samples of summer and autumn.Moreover,the habitats of samples are also related to IR spectra to some certain extent.Conclusion: The harvest seasons of Herba Sedi from Hubei Province can be preliminarily ascertained according to the IR spectra features.IR technique should be paid attention to in research and identifi cation of the harvest periods of crude plant drugs.

Objective To investigate the effects of radiation on the apoptosis rate and expression of 7 apoptosis-related genes in well-differentiated human nasopharyngeal carcinoma cell line CNE-1.Methods CNE-1 cells were cultured in vitro.The apoptosis rates of CNE-1 cells under 0-,2-,4-,6-,and 8-Gy radiation were measured by flow cytometry.The mRNA expression levels of Bcl-2,Bcl-xl,Bcl-w,Bax,Bak,Bad,and Bid were measured by RT-PCR.The Pearson test was used for analyzing the correlation of mRNA expression with apoptosis rate and radiation dose and the correlation between apoptosis rate and survival fraction.Results The early apoptosis rate of CNE-1 cells increased gradually as the radiation dose ranged from 0 to 6 Gy,but decreased when the radiation dose was 8 Gy; the late apoptosis rate of CNE-1 cells increased as the radiation dose ranged from 0 to 8 Gy.The mRNA expression of Bax was upregulated as CNE-1 cells were irradiated,and it was positively correlated with the early/late apoptosis rate and radiation dose (P =0.000 for all comparisons).The mRNA expression of Bcl-xl was downregulated,and it was negatively correlated with the early/late apoptosis rate (P =0.005 and 0.039) ; but it showed no correlation with radiation dose (P =0.369).The mRNA expression of Bcl-2 was upregulated and reached the peak level when the radiation dose was 4 Gy,and then it fell as the radiation dose increased.The mRNA expression of Bcl-w,Bcl-2,Bad,and Bid were not correlated with the early/late apoptosis rate (P =0.058 -0.894).There was no correlation between the apoptosis rate and survival fraction in CNE-1 cells (P =0.064).Conclusions Bax and Bcl-xl have some correlation with apoptosis rate in CNE-1 cells,but no correlation between the apoptosis rate and survival fraction was observed.

Objective To study the efficacy and safety of low-dose azithromycin with tiotropium bromide in the treatment of stable chronic obstructive pulmonary disease.Methods A hundred and ten patients were randomized into three groups:tiotropium bromide group (36 cases,group A),azithromycin with tiotropium bromide group(38 cases,group B)and control group(36 cases,group C).Patients in group A were given tiotropium bromide (18 μg,q,d )in addition to conventional treatment.The patients in group B were given lowdose azithromycin (250 mg,twice a week) in combined with tiotropium bromide.The patients in control group were given the conventional treatment only.The courses of treatment lasted for six months.Results Compared with the control group,the frequency of acute exacerbation in patients treated with azithromycin and tiotropium bromide was reduced remarkably ( 2.1 ± 0.6 and 4.9 ± 0.7,t =18.5061,P ＜ 0.05 ).The severity of clinic symptoms ( Cough 1.3 ± 0.5 vs.2.2 ± 0.6,P ＜ 0.05 ),expectoration ( 1.0 ± 0.2 vs.1.7 ± 0.3,P ＜ 0.05 ),anhelation ( 1.5 ± 0.8 vs.2.1 ± 0.6,t =3.6342,P ＜ 0.001 ) ],life quality ( 29 ± 8 vs.42 ± 11,P ＜ 0.05 ) and six-minutes walking distance( [ 370.00 ± 14.26 ] m vs.[ 290.00 ± 12.85 ] m,P ＜ 0.05 ) of the azithromycin with tiotropium bromide group were improved significantly when compared with control.Compared with the tiotropium bromide group,the frequency of acute exacerbation ( 2.1 ± 0.6 vs.3.2 ± 0.8,P ＜ 0.05 ),the severity of clinic symptoms (Cough 1.3 ±0.5 vs.1.8 ±0.4,P＜0.05),expectoration( 1.0 ±0.2 vs.1.3 ±0.3,P ＜0.05) and anhelation( 1.5 ±0.8 vs.1.9 ± 0.6,P ＜ 0.05 ) ],life quality ( 29 ± 8 vs.36 ± 10,P ＜ 0.05 ) and six-minutes walking distance ( [ 370.00 ± 14.26 ] m vs.[ 330.00 ± 13.76 ] m,P ＜ 0.05 ) were improved over those of tiotropium bromide group.Conclusion The long-term low-dose azithromycin in combinned with tiotropium bromide is good and safe in treating stable COPD.Therefore,it is worth of further clinical evaluation.

Objective To explore the relationship between condition of hypoxia and prognosis in patient with renal clear cell carcinoma (RCCC).Methods The expression of hypoxia-inducible factor-lo( HIF-1 α) protein in cancer tissue of 89 RCCC cases was examined by streptavidin-biotin complex immunohistochemistry.Clinical and pathological data and prognosis of 89 cases were analyzed retrospectively.There were 66 males and 23 females,with an average age of 57 years.The patients were divided into two groups,the chronic pulmonary disease (CPD) group ( 19 cases) and non CPD (NCPD) group (70 cases),with 46cases in clinical stage I,15 cases in stage Ⅱ,26 cases in stage Ⅲ,and 2 cases in stage Ⅳ.The relationship between survival time and clinicopathological variables including the presence of CPD,the positive rate of HIF-1α protein,smoking history and hemoglobin level were evaluated by the Kaplan-Meier method.And the Cox proportional hazards regression model was build to analyze the correlation between each variable and survival time.Results The 89 cases were followed up for a median time of 19 months (6 to 84 months).Twenty cases died,and 69 cases survived.Between the CPD group and NCPD group,clinical stage,hemoglobin level and the degree of expression of HIF-1 α were significantly different (P ＜ 0.05 ).The median survival time was 44 and 71 months in CPD group and NCPD group,respectively,and the difference was significant ( P ＜ 0.05 ).The median survival time was 43 and 70 months in Hb≤ 110 g/L group and Hb ＞ 110 g/L group,respectively,and the difference was significant ( P ＜ 0.05).The stronger the degree of expression of HIF-lα was,the shorter the overall survival was.And the difference was significant ( P ＜0.05 ).Associated with CPD,hemoglobin level,the expression of H1F-1α were independent factors affecting overall survival of the patients (P ＜0.05 ).CPD and HIF-1 α expression were positively related to disease-free survival time,and hemoglobin level was negatively related to disease-free survival time.Conclusions Systemic hypoxia caused by CPD may aggravate the hypoxie state in the organization of the patients with RCCC.The condition of hypoxia and prognosis in patient with RCCC is negatively correlated.

BACKGROUND: The study of isolation, purification, culture, cell labeling, inducing factors, effects of gene transfection on cytobiology, cell carrier construction, and time window for back transplantation of cell compound pertaining to marrow stromal cells (MSCs) is still in its infancy. OBJECTIVE: To search for an in vitro culture method that can be simply and effectively obtained. DESIGN, TIME AND SETTING: The present cytological in vitro experiment was performed at the Beijing Institute of Genome, Chinese Academy of Sciences between June 2006 and July 2007. MATERIALS: Eight specific pathogen-free New Zealand rabbits, aged 6 weeks, were provided by the Laboratory Animal Center, Institute of Genetics and Development, Chinese Academy of Sciences. METHODS: Under sterile condition, 1 mL rabbit bone marrow was taken and diluted with D-Hanks solution. Following centrifugation and subsequent supernatant removal, bone marrow was re-suspended using dulbecco's modified eagle's medium (DMEM) for single cell suspension. Next, single cell suspension was dropped onto the liquid surface of equal-volume lymphocyte separation medium (density: 1.077). Subsequent to centrifugation, cloudlike mononuclear cell layer was collected and re-suspended with DMEM containing 20% fetal bovine serum. The cells were inoculated at lxl0/cm2 and purified by adherent method. When 70%-80% of flask bottom was covered, cell digestion and passage was performed.MAIN OUTCOME MEASURES: Cell growth was observed with an operating microscope. Surviving cells were counted by Trypan blue viability test. Cell identification was performed by hematoxylin-eosin staining. Through the use of 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay, cell viability was detected to observe the cellular resuscitation of the cultured cells following cryopreservation. RESULTS: Twenty-four hours after inoculation, cells began to adhere to the wall, exhibiting short shuttle- or triangle-shaped appearance with different sizes of cellular processes. Three days later, adherent cells began to divide, and cell clusters could be found in some areas; One week later, most of cells exhibited scattered fibroblast-like growth; After passage, cells were evenly distributed with long shuttle-shaped appearance and arranged orderly. Following 3 passages, there wound be 5×106-1×107 adherent cells in 1 mL MSC suspension. Approximately 90% of MSCs survived and identified as mononuclear cells. Cells vigorously grew at days 1-6 after inoculation and reached a peak level at day 8, followed by a viability decline. After 56 days of resuscitation, frozen cells displayed a good and rapid growth. CONCLUSION: Highly purified MSCs can be acquired by gradient centrifugation of lymphocyte separation medium. Enough seeded cells can be obtained by in vitro culture and the cellular viability does not change after frozen preservation and resuscitation.

BACKGROUND: SOX-9 plays an important role in occurrence and development of cartilaginous tissues,enhances agglutination of mesenchymal cells,has structural domains of transcriptional activation,and can directly regulate the transcription,OBJECTIVE: To construct pDC316-SOX-9 plasmid for transfection of rabbit bone marrow stromal cells (BMSCs) using SOX-9 gene,and to investigate the effects of SOX-9 gene on growth characteristics of BMSCs and product expression. DESIGN,TIME AND SETTING: The cell gene engineering in vitro experiment was performe,d at the Beijing Institute of Genome, Chinese Academy of Scienees from June 2006 to January 2007.MATERIALS: Eight SPF New Zealand rabbits aged 6 weeks were used for culture of BMSCs.METHODS: pDC316-SOX-9 plasmid was used for transfection of rabbit BMSCs by liposome mediated method,Cell transfection included a SOX-9 plasmid transfection group,a blank plasmid group and a blank control group (only treatment of liposome). MAIN OUTCOME MEASURES: Cell morphology; growth activity; The SOX-9 protein expression in rabbit BMSCs were detected by immunohistochemistry,hematoxylin-eosin staining,reserve transcriptase-polymcrase chain reaction (RT-PCR) and enzyme-labeled immunosorbent assay (ELISA). RESULTS: Some cell colonies were detected at 4 days after pDC316-SOX-9 plasmid transfection.Spindle-shaped cells were collected after clone amplification.Cells in the blank control group gradually died over time.Cell activities in the SOX-9 plasmid transfeetion group and the blank plasmid group significantly prolonged,reached a peak at 2 weeks of transfection,and then gradually decreased.At 6 days,BMSCs were yellow in the SOX-9 plasmid transfection group following immunohistochemistry,expressing SOX-9 protein.Hematoxyliu-eosin staining showed many dikaryocytes,rich cell proliferation,similar to chondroblasts.No SOX-9 protein expression and unproductive cell proliferation in the blank plasmid group.SOX-9 mRNA was detected in the SOX-9 plasmid transfection group,but SOX-9 mRNA was not detected in the blank plasmid group and blank control group.SOX-9 levels were significantly higher in the SOX-9 plasmid transfection group than in the blank plasmid group and blank control group at 24,48 and 72 hours,1,2 weeks (P＜ 0.01).CONCLUSION: Rabbit BMSCs were successfully transfected with pDC316-SOX-9 plasmid to enhance cell growth activity and to persistently stably secrete SOX-9 protein,resulting in the differentiation of BMSCs into cartilages.

BACKGROUND: Interleukin-6 (IL-6), osteocalcin (OC) and bone alkaline phosphatase (BALP) are involved in bone metabolism, which is correlative with osteoporosis.OBJECTIVE: To observe expression properties of IL-6, OC and BALP in rats with osteoporosis.DESIGN: Control experiment with observation of randomized group division was designed.SETTING: Orthopedics Department of Shenzhen People's Hospital (2nd Affiliated Hospital of Medical College of Jinan University)MATERIALS: The experiment was performed in Animal Laboratory Room and Center Laboratory Room of Shenzhen People's Hospital from January 2002 to January 2003, in which, 60 SD female rats of 6 months were employed, averagely weighted 250 g. They were randomized into 3 groups, named ovariectomized (OVT) group, control and blank group, 20 rats in each.METHODS: In OVT group, the ovaries of rat were removed bilaterally to establish osteoporosis model. In the control, after the ovary was lifted out of abdominal cavity when the abdomen was opened, it was returned back and the abdomen was closed. In blank group, no any management was performed. In 2, 3, 4, 5 and 6 months after surgery, 4 rats were randomized from each group for assays of bone density, OC, IL-6 and BALP.MAIN OUTCOME MEASURES: Changes of bone density, OC, IL-6 and BALP of whole body of rat.assay in rats: In 4, 5 and 6 months, in OVT group, bone density was reduced remarkably compared with the control and blank group [in 4 months:(0.139 5±0.007 8), (0.147 0±0.000 8), (0.145 9±0.002 9) g/cm2; in 5 months: (0.137 9±0.000 9), (0.145 6±0.000 8), (0.144 7±0.000 5) g/cm2; in 6 months: (0.122 6±0.000 4), (0.145 0±0.002 1), (0.144 0±0.000 9) g/cm2, P sults of IL-6 assay in rats: At every time spots, IL-6 in OVT group was sults of BALP in serum of rats: In 4, 5 and 6 months, BALP in OVT group was higher significantly than the control and blank group [(2 026±4) vs (1 247±12), (1 291±7) nkat/L, (2 342±9) vs (1 273±18), (1 342±12) nkat/L,(2 633±15) vs (1 340±9), (1 357±8) nkat/L, P ＜ 0.05].CONCLUSION: In the rats with osteoporosis, bone density is decreased significantly and IL-6, OC and BALP are significantly expressed, which can be taken as the indexes for diagnosis and screen of osteoporosis.Zhang XM, Xu ZS, Xiao DM, Li R.Expression properties of interteukin-6, osteocalcin and bone alkaline phosphatase in osteoporosis rats Zhongguo Linchuang Kangfu 2005;9(43):190-2(China) [www.zglckf.com]INTRODUCTIONOsteoporosis induces complications, like bone fracture, which is associated with various factors, such as estrogen level. Osteocalcin (OC) is a kind of active multi-peptide secreted from osteoblasts,which plays the importance in regulating calcium metabolism and is a new biochemical mark in study on bone metabolism. Interleukin-6 (IL-6) provides powerful bone absorption and stimulation,which plays the important role in coupled information exchange ofbone absorption and formation. In this experiment, ovarietomy was done bilaterally in SD female rats to establish osteoporosis, afterwards, bone density, serum OC, IL-6 and bone alkaline phosphatase (BALP) were determined to probe into their relationship with the incidence of osteoporosis.

Objective To investigate the effect of sevoflurane postconditioning on the myocardial oxidative stress injury in patients undergoing heart valve replacement with cardiopulmonary bypass (CPB) . Methods Thirty ASA Ⅱ or Ⅲ and NYHA class Ⅱ or ID patients, aged 30-59 yr, weighing 42-62 kg, scheduled for cardiac valve replacement with CPB, were randomly divided into 2 groups ( n = 15 each) : control group (group C) and sevoflurane postconditioning group (group S) . Anesthesia was induced with iv injection of midazolam 0.05-0.08 mg/kg, fentanyl 3-6 μg/kg, vecuronium 0.10-0.15 mg/kg and etomidate 0.1-0.2 mg/kg. The patients were tracheal intu- bated and mechanically ventilated. Anesthesia was maintained with intermittent iv boluses of fentanyl and midazolam and continuous infusion of atracurium and propofol. In group S, 2% sevoflurane was given over 15 min via the cardiopulmonary bypass machine immediately after aortic unclamping. Blood samples from the internal jugular vein were collected immediately before skin incision (T1 ) and at 30 min, 3 h and 24 h after aortic unclamping (T2-4 ) for measurement of the plasma malondialdehyde level. Myocardial tissues were taken from the left auricle before operation and after termination of CPB for determination of α-glutathione-S-transferase expression by Western blot. Results The plasma malondialdehyde concentration was significantly lower at T2, 3, while a-glutathione-S-transferase expression in myocardial tissues higher after termination of CPB in group S than in group C ( P ＜ 0.05) . Conclusion Sevoflurane postconditioning can enhance the antioxidant capacity and attenuate the myocardial oxidative stress injury in patients undergoing cardiac valve replacement with CPB, which may be helpful to reduce myocardial ischemia-reperfusion injury.

Objective To establish a C57BL/6 mouse model of intestinal infection induced by S.Typhimurium.Methods In order to improve the infectious sensitivity of S.Typhimurium, C57BL/6 mice were intragastrically given 5% (w/v) NaHCO3.Then mice were challenged with S.Typhimurium.The health condition, survival and body weight of mice were observed from day 0 to day 7 after the bacterial infection.The pathological changes were also examined.Results the mice challenged with S.Typhimurium showed decreased body weight and typical clinical signs, including in appetence, piloerection and low survival rate.Macroscopic dissection revealed that intestinal hyperemia and swelling were founded in the mice challenged with S.Typhimurium.Histopathology showed intestinal epithelial and mucosal damages.Conclusions We have successfully established a C57BL/6 mouse model of S.Typhimurium infection.This model may be of crucial significance for studying the biological functions of associated immunological molecules or cytokines in the process of inflammatory bowel disease induced by S.Typhimurium.